Advantages of formate dehydrogenase reaction for efficient NAD+ quantification in biological samples.
Anal Biochem
; 603: 113797, 2020 08 15.
Article
em En
| MEDLINE
| ID: mdl-32562604
ABSTRACT
The medical significance of NAD+-dependent metabolic regulation acquires increasing attention, demanding rapid and clinically feasible quantification of NAD+ in complex biological samples. Here we describe the usage of formate dehydrogenase for a straightforward and highly specific fluorometric assay of NAD+ in tissue extracts, not requiring chromatographic separation of nucleotides. The assay employs the irreversible reaction of formate oxidation coupled to NAD+ reduction, catalyzed by the enzyme which has high affinity and specificity to NAD+, and is stable under a variety of conditions. The assay reliably quantifies NAD+ in the methanol extracts of the rat brain cortex and mitochondria.
Palavras-chave
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Fluorometria
/
Formiato Desidrogenases
/
NAD
Idioma:
En
Ano de publicação:
2020
Tipo de documento:
Article